High-resolution 1H NMR spectroscopic investigation of a chick embryo model of neural tube development.

High-field (1)H nuclear magnetic resonance (NMR) spectroscopy together with cryogenic probe capabilities have been applied to obtain metabolic profiles of chick embryos and to determine the suitability of the platform for profiling such mass-limited samples. Metabolic profiles were generated for both pooled and single embryo samples at early stages of neural development, using both 600 and 800 MHz (1)H NMR spectrometer platforms. High-resolution metabolic profiles, representing metabolites from many chemical classes, including triglycerides, organic acids, carbohydrates, amino acids and nucleosides, were rapidly acquired. Neural tube defects (NTDs) are severe congenital malformations, and evidence exists for prevention of NTDs by periconceptional supplementation of the diet with folate. The molecular basis for the protective ability of folate in prevention of NTDs is not clear, although the involvement of methylation has been postulated. Thus, the metabolic profiles of chick embryos following inhibition of enzymes of the methylation cycle and the effect of their action on neural tube closure were investigated. The embryos were profiled at early stages of development, and closure of the neural tube was followed via digital imaging. Metabolic profiles of embryo samples representing both neural tube closure and the neural tube remaining open were discriminated; glucose levels were found to be significantly higher in methylation-inhibited samples. The application of a non-targeted metabolic profiling approach for the study of a chick embryo model of NTDs is novel and presents the exciting potential to provide metabolic insight necessary to elucidate the complex interplay of one-carbon moiety metabolism and NTDs.